A kind of method and special-purpose dna bar code fragment of the identification method of Pleurotus eryngii strain

A barcode, eryngium eryngii technology, applied in DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of unreliable results and time-consuming, improve the identification accuracy and shorten the identification time Effect

Active Publication Date: 2018-02-06
LUDONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods are time-consuming, have certain tissue and developmental stage specificity, and the results are not reliable

Method used

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  • A kind of method and special-purpose dna bar code fragment of the identification method of Pleurotus eryngii strain
  • A kind of method and special-purpose dna bar code fragment of the identification method of Pleurotus eryngii strain
  • A kind of method and special-purpose dna bar code fragment of the identification method of Pleurotus eryngii strain

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1, the establishment of the acquisition of DNA barcode fragment and method for identification of Pleurotus eryngii strains

[0038] 1. Genomic DNA acquisition of Pleurotus eryngii strains

[0039] Genomic DNA of Pleurotus eryngii strains was extracted.

[0040] 2. PCR amplification

[0041] Using the genomic DNA obtained in 1 above as a template, PCR amplification was performed with primer 728F and primer 1567R.

[0042] 728F: 5'-CATCGAGAAGTTCGAGAAGG-3' (SEQ ID NO: 1)

[0043] 1567R: 5'-ACHGTRCCRATACCACCRATCTT-3' (SEQ ID NO: 2)

[0044] The above-mentioned PCR amplification system is as follows in Table 1:

[0045] Table 1 is the composition of the PCR reaction system (25 μL)

[0046]

[0047] The above PCR amplification program is: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 65°C for 55 s, decreasing 1°C for each cycle, extension at 72°C for 90 s, 10 cycles; denaturation at 94°C for 30 s, annealing at 55°C for 55 s...

Embodiment 2

[0053] Embodiment 2, distinguish Pleurotus eryngii bacterial classification

[0054] 1. Specific detection by the method of the present invention

[0055] The genomic DNAs of known varieties of Pleurotus eryngii strains and Pleurotus abalonus strains were extracted respectively.

[0056] Using the genomic DNA of each variety as a template, the above-mentioned primers 728F and 1567R were used to amplify to obtain PCR products of each variety.

[0057] The PCR product of each kind is sent for sequencing, and the results are as follows:

[0058] The nucleotide sequence of the PCR product of the known varieties of Pleurotus eryngii strains is sequence 3 (DNA barcode fragment for identification of Pleurotus eryngii).

[0059] The nucleotide sequence of the PCR product of known varieties of oyster mushroom strains is sequence 4 (the 1st-20th position of the sequence 4 is the upstream primer 728F, and the 385th-407th position of the sequence 4 is the reverse complementary sequence ...

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Abstract

The invention discloses a method for identifying pleurotus eryngii species and a special DNA barcode fragment. The invention provides a DNA fragment as follows: 1) a DNA fragment shown by the sequence 3; or 2) a fragment with over 99% of homology with the nucleotide sequence of the fragment in 1). The pleurotus eryngii DNA barcode fragment provided by the invention can be used for identifying the pleurotus eryngii species; by adopting the fragment for identification, the pleurotus eryngii species identifying time can be shortened while the identifying accuracy is improved; and by adopting the method, the pleurotus eryngii species can be accurately identified in 48 hours at the soonest, and the consequence that the species is discovered wrong after cultivation and fruiting is avoided.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for identifying Pleurotus eryngii strains and a special DNA barcode fragment. Background technique [0002] Since the reform and opening up, my country's edible fungi industry has developed rapidly and has become the world's largest producer of edible fungi. Pleurotus eryngii is a new species of rare edible fungus that has been successfully cultivated in recent years and integrates edible, medicinal and dietotherapy. Pleurotus eryngii is rich in nutrition, rich in protein, carbohydrates, vitamins and minerals such as calcium, magnesium, copper, zinc, etc. Strains play an important and irreplaceable role in the development of Pleurotus eryngii industry. In recent years, most of the strain accidents are caused by wrong strains. Unscrupulous practitioners use fake and shoddy strains to disrupt the market and seek improper benefits. Therefore, the authenticity detection of Pl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/04C12N15/11
CPCC12Q1/6895
Inventor 赵鹏纪森鹏周军辉高兴喜
Owner LUDONG UNIVERSITY
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